中华皮肤科杂志 ›› 2012, Vol. 45 ›› Issue (7): 488-491.

• 论著 • 上一篇    下一篇

银屑病患者骨髓基质细胞分泌转化生长因子β1、干细胞因子、角质形成细胞生长因子、肿瘤坏死因子α水平检测

潘智慧1,王丽2,贾珍3,刘瑞风4,王荣4,张永翠5,李欣4,张开明4   

  1. 1. 山西大医院皮肤科
    2. 山西医科大学第二医院皮肤科
    3. 山西医科大第二医院
    4. 太原市中心医院皮肤科
    5. 太原市中心医院
  • 收稿日期:2011-09-26 修回日期:2012-03-06 出版日期:2012-07-15 发布日期:2012-07-02
  • 通讯作者: 王丽 E-mail:lys0633@163.com
  • 基金资助:

    国家自然科学基金资助项目

Detection of transforming growth factor β1, stem cell factor, keratinocyte growth factor and tumor necrosis factor-α secreted by bone marrow stromal cells from patients with psoriasis vulgaris

  • Received:2011-09-26 Revised:2012-03-06 Online:2012-07-15 Published:2012-07-02
  • Contact: Wang Li E-mail:lys0633@163.com

PDF

18

赞

摘要：

目的 通过检测银屑病患者骨髓基质细胞分泌转化生长因子（TGF-β1）、干细胞因子（SCF）、肿瘤坏死因子α（TNF-α）和角质形成细胞生长因子（KGF）水平，探讨银屑病患者骨髓造血微环境的变化。方法 寻常性银屑病患者20例，进行期10例，静止期10例，银屑病皮疹面积和严重程度（PASI）范围为 0.6 ～ 22.8，平均10.97。健康对照组20例为骨髓象正常者。采用密度梯度离心法分离骨髓单一核细胞，通过贴壁法培养骨髓基质细胞。收集传3代后继续培养72 h的骨髓基质细胞及上清液，用流式细胞仪鉴定细胞表面标志，ELISA法检测上清液中TNF-α、TGF-β1、KGF和SCF的表达水平。银屑病组和对照组间各检测指标比较采用t检验，细胞因子与PASI相关性分析采用Pearson相关分析。结果 倒置显微镜下，银屑病组与健康对照组的骨髓基质细胞形态无显著差异，90%以上表面抗原CD29高表达，而CD34、CD45及HLA-DR表达为阴性。银屑病患者骨髓基质细胞分泌的TNF-α（22.93 ± 10.11 μg/L）显著低于健康对照组（35.73 ± 15.15 μg/L，两组比较，t = 3.14，P ＜ 0.05），SCF 表达水平（76.80 ± 16.19 μg/L）明显高于健康对照组（59.86 ± 22.41 μg/L，两组比较，t = 2.74，P ＜ 0.05），而KGF 、TGF-β1 的分泌量与健康对照组比较差异均无统计学意义（P ＞ 0.05）。银屑病患者组SCF、TNF-α、KGF、TGF-β1 分泌水平与PASI无相关关系（P值均 ＞ 0.05）。结论 银屑病患者骨髓基质细胞分泌的TNF-α、SCF存在异常，表明银屑病患者骨髓造血微环境存在异常。

关键词: 细胞因子

Abstract:

Objective To assess the changes in bone marrow microenvironment in patients with psoriasis by determining the level of transforming growth factor β1 （TGF-β1）, stem cell factor （SCF）, keratinocyte growth factor （KGF） and tumor necrosis factor-α （TNF-α） secreted by bone marrow stromal cells（BMSCs）. Methods This study recruited 20 healthy controls with normal bone marrow picture and 20 patients with psoriasis vulgaris, including 10 at progressive stage and 10 at resting stage. The psoriasis area and severity index （PASI） score varied from 0.6 to 22.8 and averaged 10.97 in these patients. Bone marrow mononuclear cells were isolated by density-gradient centrifugation from bone marrow of these subjects, and BMSCs were cultured with adherent method. After three passages, the BMSCs were subjected to a 72-hour culture followed by the identification of cell phenotypes via flow cytometry and determination of TGF-β1, SCF, KGF and TNF-α levels in the culture supernatant via enzyme linked immunosorbent assay （ELISA）. The parameters were compared by two independent samples t test between the two groups, and the correlation of cytokines with PASI was assessed by Pearson correlation analysis. Results Inverted microscopy revealed no obvious difference in the morphology of BMSCs between the patients and controls. CD29 was expressed by more than 90% of the BMSCs, but no expression of CD45, CD34 or HLA-DR was observed in them. The BMSCs from patients showed a significantly lower level of supernatant TNF-α （（22.93 ± 10.11 ） μg/L vs. （35.73 ± 15.15） μg/L， t = 3.14，P ＜ 0.05）, a higher level of supernatant SCF （（76.80 ± 16.19） μg/L vs. （59.86 ± 22.41） μg/L, t = 2.74, P ＜ 0.05）, and a similar level of supernatant KGF and TGF-β1 （both P > 0.05） compared with those from the controls. The PASI score was uncorrelated with the levels of SCF, TNF-α, KGF or TGF-β1 secreted by BMSCs in patients with psoriasis （all P > 0.05）. Conclusions The levels of SCF and TNF-α secreted by BMSCs are aberrant in patients with psoriasis, hinting an abnormal bone marrow microenvironment in these patients.

Key words: cell factors